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Generation of a vector suite for protein solubility screening

Recombinant protein expression has become an invaluable tool for academic and biotechnological projects. With the use of high-throughput screening technologies for soluble protein production, uncountable target proteins have been produced in a soluble and homogeneous state enabling the realization o...

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Autores principales: Correa, Agustín, Ortega, Claudia, Obal, Gonzalo, Alzari, Pedro, Vincentelli, Renaud, Oppezzo, Pablo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3934309/
https://www.ncbi.nlm.nih.gov/pubmed/24616717
http://dx.doi.org/10.3389/fmicb.2014.00067
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author Correa, Agustín
Ortega, Claudia
Obal, Gonzalo
Alzari, Pedro
Vincentelli, Renaud
Oppezzo, Pablo
author_facet Correa, Agustín
Ortega, Claudia
Obal, Gonzalo
Alzari, Pedro
Vincentelli, Renaud
Oppezzo, Pablo
author_sort Correa, Agustín
collection PubMed
description Recombinant protein expression has become an invaluable tool for academic and biotechnological projects. With the use of high-throughput screening technologies for soluble protein production, uncountable target proteins have been produced in a soluble and homogeneous state enabling the realization of further studies. Evaluation of hundreds conditions requires the use of high-throughput cloning and screening methods. Here we describe a new versatile vector suite dedicated to the expression improvement of recombinant proteins (RP) with solubility problems. This vector suite allows the parallel cloning of the same PCR product into the 12 different expression vectors evaluating protein expression under different promoter strength, different fusion tags as well as different solubility enhancer proteins. Additionally, we propose the use of a new fusion protein which appears to be a useful solubility enhancer. Above all we propose in this work an economic and useful vector suite to fast track the solubility of different RP. We also propose a new solubility enhancer protein that can be included in the evaluation of the expression of RP that are insoluble in classical expression conditions.
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spelling pubmed-39343092014-03-10 Generation of a vector suite for protein solubility screening Correa, Agustín Ortega, Claudia Obal, Gonzalo Alzari, Pedro Vincentelli, Renaud Oppezzo, Pablo Front Microbiol Microbiology Recombinant protein expression has become an invaluable tool for academic and biotechnological projects. With the use of high-throughput screening technologies for soluble protein production, uncountable target proteins have been produced in a soluble and homogeneous state enabling the realization of further studies. Evaluation of hundreds conditions requires the use of high-throughput cloning and screening methods. Here we describe a new versatile vector suite dedicated to the expression improvement of recombinant proteins (RP) with solubility problems. This vector suite allows the parallel cloning of the same PCR product into the 12 different expression vectors evaluating protein expression under different promoter strength, different fusion tags as well as different solubility enhancer proteins. Additionally, we propose the use of a new fusion protein which appears to be a useful solubility enhancer. Above all we propose in this work an economic and useful vector suite to fast track the solubility of different RP. We also propose a new solubility enhancer protein that can be included in the evaluation of the expression of RP that are insoluble in classical expression conditions. Frontiers Media S.A. 2014-02-25 /pmc/articles/PMC3934309/ /pubmed/24616717 http://dx.doi.org/10.3389/fmicb.2014.00067 Text en Copyright © 2014 Correa, Ortega, Obal, Alzari, Vincentelli and Oppezzo. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Correa, Agustín
Ortega, Claudia
Obal, Gonzalo
Alzari, Pedro
Vincentelli, Renaud
Oppezzo, Pablo
Generation of a vector suite for protein solubility screening
title Generation of a vector suite for protein solubility screening
title_full Generation of a vector suite for protein solubility screening
title_fullStr Generation of a vector suite for protein solubility screening
title_full_unstemmed Generation of a vector suite for protein solubility screening
title_short Generation of a vector suite for protein solubility screening
title_sort generation of a vector suite for protein solubility screening
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3934309/
https://www.ncbi.nlm.nih.gov/pubmed/24616717
http://dx.doi.org/10.3389/fmicb.2014.00067
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